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【答案】应助回帖
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huojinlong8610: 金币+500, 翻译EPI+1, ★★★★★最佳答案, 不错，速度快，翻译的行 2012-04-13 00:55:55
Mally89: 金币+3, 感谢应助！~欢迎常来！·好运！~\(^o^)/~ 2012-04-14 13:39:16
BMI is the use of our unique resources of native pig breeds using strict scientific breeding methods to select consecutive height of nearly Cross, to cultivate the success of the world's first large-scale mammalian laboratory animals inbred for bio-medicine, xenotransplantation research large inbred experimental animals. Over the years we have been committed to the BMI in the basis of genetic breeding work, but the genetic level to explore the genetic features of a relatively small, especially the genetic characteristics of genes related to the level of the important features of its growth, metabolism, immune have not been systematic research.
Further insight into the genetic basis of the molecular level, to reflect its genetic characteristics, this study uses modern molecular biology techniques to start with the RNA or DNA, the level of gene structure and expression of BMI has not been studied with its growth and development, metabolism, immune regulation, transplant rejection, pathophysiology, breeding for disease resistance, and sex identification of germplasm genetic characteristics of 13 candidate functional genes in-depth study. These genes include: growth hormone (GH), aquaporin 3 (of AQP3 in), vascular endothelial growth factor (VEGF), tumor necrosis factor alpha (TNF-alpha), alpha interferon (IFN-alpha), interleukin-6 ( IL-6), the Y chromosome sex-determining gene (the SRY), the Fas apoptosis inhibitory molecule (FAIM1), the polyglutamine binding protein 1 (PQBP1), synaptotagmin protein 6 (STXBP6) ribosylation enzymes (RBKS ), fat acetyl-CoA reductase (FAR1), centromere protein of P (CENPP). Access to the complete coding sequence of these genes and these genes carry out a comparative genomics, bioinformatics analysis, according to the different genetic characteristics of each gene, targeted multi-organization sample fluorescence quantitative (qPCR) expression analysis, and a variety of physical and chemical characteristics and function prediction and analysis of the gene encoding the protein. In addition to the GH and the SRY gene in the prokaryotic expression analysis, and GH gene by Western Blot verification.
The results show that the relatively conservative amino acid sequence of each gene among different species, similar to high, but can also be found BMI is different from other breeds of nucleotide and amino acid level of SNP, such as found in the BMI of GH gene coding region and miniature pigs of the domestic and foreign large-scale pig and wild boar difference c.26T> C SNP, but also to prove the GH gene complete coding sequence can not be induced to express the protein, only to remove the mature peptide to protein expression. AQP3 amino acids in pigs between 100% conservative, but found a c.204C> A SNP site in the nucleotide coding region. VEGF amino acid exists a p.A102T SNP nucleotide to exist c.15G> C and c.438G> A, two SNP loci. SRY gene screening to a pair of specific primers can be a BMI of early embryonic sex identification, and found the concentration of 0.5 mM IPTG can induce a large number of SRY protein to produce. Multi-organization of the housekeeping gene 18 S correction qPCR analysis showed that the differentially expressed genes in each tissue phenomenon. The physical and chemical properties and structure of the protein molecular weight, isoelectric point, secondary structure, tertiary structure, transmembrane helices and signal peptide reflects the function and status of the BMI of each gene in the protein expression level. 13 genes have been reached with the GenBank database, authentication, registration number (Accession Number), respectively: JQ177096 (GH), HQ888860 (of AQP3 in), JF831364 (VEGF), JF831365 (TNF-alpha), JQ839262 (IFN-alpha), JQ839263 ( IL-6), GU991615 (SRY), Nm_001245976 (FAIM1), Nm_001245976 (PQBP1), Nm_001246666 (STXBP6), JF944892 (RBKS), Nm_001252425 (FAR1), JF944894 (CENPP), which FAIM1, PQBP1, STXBP6 and FAR1 genes have been GenBank database to determine the standard sequence (format ******** Nm_) and pigs, the preliminary study of these new gene sequence annotation as well as its function can be laid for the in-depth study of the regulation of expression of these genes, functional identification the basis of molecular biology.
13 genes cloned in BMI is the first study, to fill a gap of the related genes, 13 genes have been reached with the GenBank database certification of GH, of AQP3 in, of VEGF and TNF-alpha and IFN-α, IL 6 gene coding sequence is different from other breeds of the GenBank sequence as a BMI of unique haplotypes and encoded by these genes also found that more can and other breeds with each other to distinguish SNP loci; SRY BMI unique SNP in the gene sequence does not exist, but we have developed a pair of specific primers can be used as the the BMI embryonic sex identification of molecular markers; FAIM1, PQBP1, STXBP6 RBKS, FAR1 CENPP and other six genes is the first isolated pig genes, these genes are functional genes related to immune, disease, metabolic and other life activities. In this study, results not only for the Banna minipig inbred further breeding use to provide the basis for data, methods and ideas of its unique genetic characteristics at the molecular level, but also its intellectual property protection, development and experimental animals of use to provide a scientific basis. |
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